Volume 19 Issue 3
Jun.  2006
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Article Navigation >  Biomedical and Environmental Sciences  > 2006  >  19(3): 214-218

JIN ZHANG, JIAN-MEI GAO, FENG LI, JUN HAN, LAN CHEN, BAO-YUN ZHANG, XIAO-FAN WANG, WEI ZHOU, YONG LIU, XIAO-PING DONG. Establishment of Cell Free Conversion System With Biotin-labelled Recombinant PrPsen Expressed in E. Coli[J]. Biomedical and Environmental Sciences, 2006, 19(3): 214-218.
Citation: JIN ZHANG, JIAN-MEI GAO, FENG LI, JUN HAN, LAN CHEN, BAO-YUN ZHANG, XIAO-FAN WANG, WEI ZHOU, YONG LIU, XIAO-PING DONG. Establishment of Cell Free Conversion System With Biotin-labelled Recombinant PrPsen Expressed in E. Coli[J]. Biomedical and Environmental Sciences, 2006, 19(3): 214-218.
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Establishment of Cell Free Conversion System With Biotin-labelled Recombinant PrPsen Expressed in E. Coli

  • School of Medicine, Xi'an Jiaotong University, Zhuque Street 205, Xi'an 710061, China

  • State Key Laboratory for Infectious Disease Prevention and Control, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Yingxin Rd 100, Beijing 100052, China

  • State Key Laboratory for Infectious Disease Prevention and Control, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Yingxin Rd 100, Beijing 100052, China;National Laboratory of Medical Molecular Biology, Institute of Basic Medical Science, Chinese Academy of Medical Sciences and Peking Union Medical College, Dongdan Santiao 5, Beijing 100005, China

Funds:  国家自然科学基金(30070038;30130070)%国家高技术研究发展计划(863计划)(2001AA215391)%EU Project QLRT(2000 01441)
  • Objective To report a protocol using biotin-labelled PrP protein in cell free conversion assay instead of isotope. Methods A hamster PrP protein (HaPrP) was expressed in E. coli and purified with HIS-tag affinity chromatograph. After being labelled with biotin, HaPrP was mixed with PrPSc preparation from scrapie strain 263K. Results Protease-resistant bands were detected after four-day incubation. Conclusion The new conversion model provides a reliable, easily handling, and environment-friendly method for studies of prion and transmissible spongiform encephalopathies.
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Establishment of Cell Free Conversion System With Biotin-labelled Recombinant PrPsen Expressed in E. Coli

  • School of Medicine, Xi'an Jiaotong University, Zhuque Street 205, Xi'an 710061, China
  • State Key Laboratory for Infectious Disease Prevention and Control, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Yingxin Rd 100, Beijing 100052, China
  • State Key Laboratory for Infectious Disease Prevention and Control, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Yingxin Rd 100, Beijing 100052, China;National Laboratory of Medical Molecular Biology, Institute of Basic Medical Science, Chinese Academy of Medical Sciences and Peking Union Medical College, Dongdan Santiao 5, Beijing 100005, China
Funds:  国家自然科学基金(30070038;30130070)%国家高技术研究发展计划(863计划)(2001AA215391)%EU Project QLRT(2000 01441)

Abstract: Objective To report a protocol using biotin-labelled PrP protein in cell free conversion assay instead of isotope. Methods A hamster PrP protein (HaPrP) was expressed in E. coli and purified with HIS-tag affinity chromatograph. After being labelled with biotin, HaPrP was mixed with PrPSc preparation from scrapie strain 263K. Results Protease-resistant bands were detected after four-day incubation. Conclusion The new conversion model provides a reliable, easily handling, and environment-friendly method for studies of prion and transmissible spongiform encephalopathies.

JIN ZHANG, JIAN-MEI GAO, FENG LI, JUN HAN, LAN CHEN, BAO-YUN ZHANG, XIAO-FAN WANG, WEI ZHOU, YONG LIU, XIAO-PING DONG. Establishment of Cell Free Conversion System With Biotin-labelled Recombinant PrPsen Expressed in E. Coli[J]. Biomedical and Environmental Sciences, 2006, 19(3): 214-218.
Citation: JIN ZHANG, JIAN-MEI GAO, FENG LI, JUN HAN, LAN CHEN, BAO-YUN ZHANG, XIAO-FAN WANG, WEI ZHOU, YONG LIU, XIAO-PING DONG. Establishment of Cell Free Conversion System With Biotin-labelled Recombinant PrPsen Expressed in E. Coli[J]. Biomedical and Environmental Sciences, 2006, 19(3): 214-218.
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